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[Biodegradation of polycyclic aromatic hydrocarbons and its application to the bioremediation of contaminated soils and sludges].

Polycyclic aromatic hydrocarbons (PAHs) are a group of priority pollutants due to their toxic, mutagenic, and carcinogenic properties. They are found in high concentration in many soils and sludges, mainly those associated with petroleum, gas-production and wood-preservation industries. Application of microorganisms for the remediation of contaminated soils and sludges has gained interest as a promising technique in comparison to the conventional practices of landfilling, incineration or solvent extraction. This paper reviews the controlling variables of the biodegradation of PAHs as well as the use of solid-phase and slurry-phase bioremediation techniques. The acceptance of bioremediation technology requires the demonstration of its efficacy, reliability, predictability of the field performance from lab-scale treatability studies, and economical advantage. Some limitations in the biodegradation of high-molecular-weight PAHs and failures in the control of scale-depending factors (mass transport, spatial heterogeneities, abiotic losses) can reduce its effectiveness. Methodology used for the systematic evaluation of bioremediation is also reviewed.

Biodegradation, Environmental↗

In situ biosurfactant production by Bacillus strains injected into a limestone petroleum reservoir.

Biosurfactant-mediated oil recovery may be an economic approach for recovery of significant amounts of oil entrapped in reservoirs, but evidence that biosurfactants can be produced in situ at concentrations needed to mobilize oil is lacking. We tested whether two Bacillus strains that produce lipopeptide biosurfactants can metabolize and produce their biosurfactants in an oil reservoir. Five wells that produce from the same Viola limestone formation were used. Two wells received an inoculum (a mixture of Bacillus strain RS-1 and Bacillus subtilis subsp. spizizenii NRRL B-23049) and nutrients (glucose, sodium nitrate, and trace metals), two wells received just nutrients, and one well received only formation water. Results showed in situ metabolism and biosurfactant production. The average concentration of lipopeptide biosurfactant in the produced fluids of the inoculated wells was about 90 mg/liter. This concentration is approximately nine times the minimum concentration required to mobilize entrapped oil from sandstone cores. Carbon dioxide, acetate, lactate, ethanol, and 2,3-butanediol were detected in the produced fluids of the inoculated wells. Only CO(2) and ethanol were detected in the produced fluids of the nutrient-only-treated wells. Microbiological and molecular data showed that the microorganisms injected into the formation were retrieved in the produced fluids of the inoculated wells. We provide essential data for modeling microbial oil recovery processes in situ, including growth rates (0.06 +/- 0.01 h(-1)), carbon balances (107% +/- 34%), biosurfactant production rates (0.02 +/- 0.001 h(-1)), and biosurfactant yields (0.015 +/- 0.001 mol biosurfactant/mol glucose). The data demonstrate the technical feasibility of microbial processes for oil recovery.

Bacillus↗

Influence of lipids with and without other cottonseed reserve materials on aflatoxin B(1) production by Aspergillus flavus.

Cottonseed storage lipids (primarily triglycerides), in either crude or refined form, were found to support growth and aflatoxin B(1) production by Aspergillus flavus. When lipids were removed from ground whole cottonseed by petroleum ether extraction, aflatoxin production dropped by more than 800-fold. Reconstitution of the lipid-extracted ground whole seed with a crude preparation of cottonseed lipids restored aflatoxin production to the previous levels. Fungal utilization of the three major cottonseed reserve materials, raffinose, triglycerides (refined cottonseed oil), and cottonseed storage protein, was monitored in vitro over a 7 day fermentation period. The fermentation medium contained the reserve compounds in proportions approximating those found in mature cottonseed. A. flavus rapidly converted raffinose to fructose and melibiose, presumably by action of invertase, and then hydrolyzed the melibiose. These simple sugars apparently supported initial growth and aflatoxin B(1) production. Raffinose and the resulting melibiose were nearly exhausted by day 2. Fungal hydrolysis of triglycerides began as exhaustion of carbohydrate approached. After day 2, rapid catabolism of the released fatty acids began and coincided with glucose regeneration through gluconeogenesis, which peaked on day 6. The fungus did not preferentially utilize specific fatty acids. A. flavus also produced a number of storage metabolites, including arabitol, erythritol, mannitol, and trehalose. Mannitol was produced in much higher concentrations than the other storage metabolites. Selective use of simple carbohydrates by A. flavus to drive aflatoxin production may suggest strategies for reducing vulnerability of cottonseed to aflatoxin contamination.

Aflatoxin B1↗

Update of a study of crude oil production workers 1946-94.

OBJECTIVE: To update information on workers in the petroleum industry engaged in the production of crude oil to determine whether the patterns of mortality have changed with 14 additional years of follow up. METHODS: All workers were employed at company production and pipeline locations sometime during 1946-94. The cohort now consists of 24 124 employees with an average of 22 years of follow up. RESULTS: The overall mortality, and most cause specific mortalities were lower than or similar to those for the general United States population. For white men (81% of the cohort), there were 4361 observed deaths and 5945 expected, resulting in a significantly lower standardised mortality ratio (SMR) of 73. There were significant deficits for all the leading causes of death in the United States including all cancers, cancer of the lung, stroke, heart disease, respiratory disease, and accidents. Slightly increased mortality was found for cancer of the prostate, cancer of the brain and central nervous system, and cancer of other lymphatic tissue. For benign and unspecified neoplasms, the SMR was 152 (95% confidence interval (95% CI) 95 to 230). There was a significant increase for acute myelogenous leukaemia that was restricted to people who were first employed before 1940 and who were employed in production and pipeline jobs for >30 years. Overall mortality patterns for non-white men and women were similar to those for white men. Mortality patterns for white men were also examined by duration of employment, time first employed, and by job group. CONCLUSIONS: The results of the updated study showed a favourable mortality experience for crude oil production workers compared with the United States population.

Cause of Death↗

Single cell protein: a promising dietary substitute.

Promises and limitations in exploitation of microorganisms for single cell protein (SCP) production have been reviewed. Various aspects discussed include global protein problem, microbes as potential sources of food and feed with its advantages and disadvantages, salient features in selection of microorganisms for SCP production, production of SCP from petroleum based hydrocarbons, renewable raw materials like agricultural and forestry residues, industrial by-products and wastes, nutritional and safety evaluation of SCP, ways of improving SCP production, economics of SCP production and the present status of SCP in India.

Bacterial Proteins↗

The formation and occurrence of polynuclear aromatic hydrocarbons associated with food.

Polynuclear aromatic hydrocarbons are common contaminants of processed food, usually at trace levels. These hydrocarbons are products of combustion and pyrolysis, and are present in petroleum and coal, and in products derived from them. Most polynuclear aromatic hydrocarbons are not carcinogenic, but some of them are, and a few are potent inducers of skin and lung tumors in mice. Their carcinogenic properties have not been fully explored, but they seem to be less potent by ingestion or inhalation, and they are known as a group to produce cancer in humans. The most effective carcinogens among them are those with 5 or 6 fused rings, and these tend to be less prevalent in mixtures than the 3- and 4-ring hydrocarbons, most of which are not carcinogenic. Sophisticated analytical methods, using solvent extraction and chromatography have been developed to detect and measure polynuclear aromatic hydrocarbons at levels of 1 in 10(9) (1 part per billion) or less, and these have been applied to the measurement of individual compounds in foods, as well as in products of combustion and pyrolysis. Wood smoke and smoked foods contain the carcinogenic benzo[a]pyrene at levels of 1 ppb, and other hydrocarbons; liquid smoke has lower levels. Crude vegetable oils have higher concentrations, but purified 'deodorized' oils have benzo[a]pyrene levels near 1 ppb. Sausages cooked over burning logs had as much as 200 ppb benzo[a]pyrene. Charcoal-broiled steaks and ground meat had benzo[a]pyrene concentrations up to 50 micrograms/kg, while less fatty pork and chicken had lower concentrations (up to 10 micrograms/kg). It was probable that the rendered fat dripped on to the hot charcoal and pyrolyzed to form quantities of polynuclear aromatic hydrocarbons, which rose with the smoke to deposit on the meat. Therefore, oven cooking or cooking with a heat source above the meat, or segregation of the meat from the smoke resulted in food containing negligible amounts of polynuclear aromatic hydrocarbons. Modifications of cookings practices accordingly would greatly reduce exposure to this group of carcinogens.

Animals↗

Analysis and stability of mercury speciation in petroleum hydrocarbons.

Raw petroleum and natural gas often contain high concentrations of mercury, which can be damaging to the metal components of production facilities, as well as to the environment. Various Hg species have different properties in terms of mobility, reactivity and bioavailability. Thus, for cost-effective decisions regarding plant design, Hg extraction, and pollution control, speciation information must be available at the production facility. In this paper, a simple, wet chemical speciation method, which provides data on Hg(o), dissolved and particulate total Hg, Hg(II), and methyl Hg is presented. The method incorporates species-specific extraction and separation procedures, followed by cold vapor atomic fluorescence spectrometry (CVAFS). For each species, detection limits of approximately 0.1 ng/g were obtained. Storage experiments in various containers showed that organo-mercury species were stable for at least 30 days in all containers except those made of polyethylene; and Hg(o) was stable in all containers except those made of stainless steel or polyethylene. Hg(II) was rapidly lost from all containers except those made of aluminum, which rapidly converted it to Hg(o), which was stable. In general, most of the total Hg in petroleum products was particulate Hg, followed by dissolved Hg(II) and Hg(o). Sub-ng/g concentrations of methyl-Hg were observed in most samples.

Journal Article↗

Automated tandem mass spectrometry by orthogonal acceleration TOF data acquisition and simultaneous magnet scanning for the characterization of petroleum mixtures.

The automated acquisition of the product ion spectra of all precursor ions in a selected mass range by using a magnetic sector/orthogonal acceleration time-of-flight (oa-TOF) tandem mass spectrometer for the characterization of complex petroleum mixtures is reported. Product ion spectra are obtained by rapid oa-TOF data acquisition and simultaneous scanning of the magnet. An analog signal generator is used for the scanning of the magnet. Slow magnet scanning rates permit the accurate profiling of precursor ion peaks and the acquisition of product ion spectra for all isobaric ion species. The ability of the instrument to perform both high- and low-energy collisional activation experiments provides access to a large number of dissociation pathways useful for the characterization of precursor ions. Examples are given that illustrate the capability of the method for the characterization of representative petroleum mixtures. The structural information obtained by the automated MS/MS experiment is used in combination with high-resolution accurate mass measurement results to characterize unknown components in a polar extract of a refinery product. The exhaustive mapping of all precursor ions in representative naphtha and middle-distillate fractions is presented. Sets of isobaric ion species are separated and their structures are identified by interpretation from first principles or by comparison with standard 70-eV EI libraries of spectra. The utility of the method increases with the complexity of the samples.

Journal Article↗

Comparative carcinogenic and mutagenic activity of coal tar and petroleum asphalt paints used in potable water supply systems.

Coal tar and petroleum asphalt paints are among the products used as coatings for water pipes and storage tanks to retard corrosion. Formulations of these coatings were tested in the Ames mutagenesis and the mouse skin carcinogenesis bioassays. To test the mutagenicity of the paints, six doses ranging from 0.005 to 10 microliters per plate were assayed. In the mouse skin bioassay, doses of the coal tar paints ranging from 0.2 to 200 microliters were administered topically to 30 SENCAR mice per group. These initiating doses were followed by applications of 1.0 micrograms of 12-o-tetradecanoyl-phorbol-13-acetate (TPA) in 0.2 ml acetone topically, three times weekly for 20 weeks. Petroleum asphalt paints were tested in groups of 40 animals at 200 and 600 microliters doses. All coal tar paints showed mutagenic activity after metabolic activation with S-9, with the highest response being in strains TA 98 and TA 100. None of the petroleum asphalt paints gave mutagenic responses. Both types of coatings resulted in positive responses in the initiation/promotion study. The coal tar paints gave rise to 1000-1800 times the tumor response observed with petroleum asphalt products. One coal tar product was positive when tested as a complete carcinogen in the mouse at 2 microliters per application once weekly for 30 weeks, whereas the asphalt paint was negative at 100 times the dose. The biological responses to the products were greater than expected from their polycyclic aromatic hydrocarbon (PAH) content. These findings suggest that the hazard posed by these coatings may not be fully explained by their PAH contents.

Animals↗

Application of high-temperature simulated distillation to the residuum oil supercritical extraction process in petroleum refining

The gas chromatographic method of high-temperature simulated distillation (HTSD) is described, and the results are presented for the application of HTSD to the characterization of petroleum refinery feed and products from solvent deasphalting operations. Results are presented for refinery residual feed, deasphalted oil, and asphaltene fractions from the residual oil supercritical extraction process. Asphaltene removal from petroleum residuum using solvent deasphalting results in the improved quality and high recovery of deasphalted oil product for use as lube oil, fluid catalytic cracking, or hydrocracker feedstocks. The HTSD procedure presented here proves valuable for characterizing the fractions from the deasphalting process to obtain the percentage yield with boiling point data over the range from approximately 36 degrees C (97 degrees F) to 733 degrees C (1352 degrees F), which covers the boiling range of n-paraffins of carbon number C5 to C108.

Journal Article↗

Application of the Salmonella mutagenicity assay and determination of polycyclic aromatic hydrocarbons in workplaces exposed to petroleum pitch and petroleum coke.

Workplaces of an Italian carbon electrode factory, exposed to petroleum pitch and petroleum coke, were studied using a coupled chemical and biological approach to evaluate occupational mutagenic/carcinogenic hazards. Analytical procedures for the determination of polycyclic aromatic hydrocarbons (PAH) and Salmonella/microsome mutagenicity tests (with TA98 and TA100 strains) were performed on both industrial ingredients (pitch and coke) and airborne particulate matter of the working environment, after fractionating by sequential Soxhlet extractions with four organic solvents of increasing polarity (benzene, chloroform, methanol and acetone). The results showed: (a) the presence of extraordinarily high PAH (carcinogenic and non-carcinogenic) contents in the benzene extracts of petroleum pitch (3.6 wt% of total PAH) and of airborne particulate samples (up to 0.35 wt% of total PAH), in correlation with very high indirect (after metabolic activation) mutagenic responses of benzene extracts with strain TA98; (b) very high indirect mutagenic responses in the other extracts of the airborne particulate samples (especially with strain TA98); (c) the production during the processing at high temperatures of directly acting mutagens (without metabolic activation) which were absent in the starting materials and their release in the air of workplaces. The comparison of chemical analytical and mutagenicity data has proved to be an interesting approach for better defining the relative health hazards due to occupational exposure to potentially mutagenic/carcinogenic petroleum products.

Air Pollutants↗

A case-control study of multiple myeloma and occupation.

Lifetime job histories from a population-based, case-control study were analyzed to investigate the relationship between multiple myeloma and employment in various occupations and industries. Interviews were obtained from 89% (692) of eligible incident cases and 83% (1683) of eligible controls. An elevated risk was observed among persons ever employed as painters [odds ratio (OR) = 2.1, 95% confidence interval (CI) = 1.2-3.6], particularly for those employed for 10 or more years (OR = 4.1, 95% CI = 1.8-10.4). A small excess risk was observed among agricultural workers employed for 10 or more years (OR = 1.3, 95% CI = 1.0-2.2), with a higher relative risk observed among farm laborers (OR = 1.8, 95% CI = 1.0-4.0). Among agricultural workers who reported having been highly exposed to pesticides, the OR was 5.2 (95% CI = 1.6-21.1). Some evidence, based on smaller numbers, was also found to support an association with firefighting and employment in the petroleum- and coal-products manufacturing industries. Little evidence was found to support the previously noted association with wood exposure, and no evidence for an association with employment in the rubber or petroleum refining industries was found. This study lends further support to previously reported associations between multiple myeloma and employment among painters and agricultural workers.

Adult↗

A review of the occurrence, toxicity, and biodegradation of condensed thiophenes found in petroleum.

Condensed thiophenes comprise a significant portion of the organosulfur compounds in petroleum and in other products from fossil fuels. Dibenzothiophene (DBT) has served as a model compound in biodegradation studies for over two decades. However, until quite recently, few other organosulfur compounds were studied, and their fates in petroleum-contaminated environments are largely unknown. This paper presents a review of the types of organosulfur compounds found in petroleum and summarizes the scant literature on toxicity studies with condensed thiophenes. Reports on the biodegradation of benzothiophene, alkylbenzothiophenes, DBT, alkylDBTs, and naphthothiophenes are reviewed with a focus on the identification of metabolites detected in laboratory cultures. In addition, recent reports on quantitative studies with DBT and naphtho[2,1-b]thiophene indicate the existence of polar sulfur-containing metabolites that have escaped detection and identification.

Biodegradation, Environmental↗

Color and chlorinated organics removal from pulp mills wastewater using activated petroleum coke.

Delayed petroleum coke, a waste by-product from the oil sand industry, was utilized in the production of activated carbon. The activated carbon was then evaluated for color and chlorinated organics reduction from pulp mill wastewater. The activation of the petroleum coke was evaluated using a fixed bed reactor involving carbonization and activation steps at temperature of 850 degrees C and using steam as the activation medium. The activation results showed that the maximum surface area of the activated coke was achieved at an activation period of 4 h. The maximum surface area occurred at burnoff and water efficiency of 48.5 and 54.3%, respectively. Increasing the activation period to 6 h resulted in a decrease in the surface area. Methylene blue adsorption results indicated that the activation process was successful. Methylene blue adsorbed per 100 g of applied activated coke was 10 times higher than that adsorbed by raw petroleum coke. Adsorption equilibrium results of the bleached wastewater and the activated coke showed that significant color, COD, DOC and AOX removal (> 90%) was achieved when the activated coke dose exceeded 15,000 mg/L. Adsorption isotherms, in terms of COD, DOC, UV and color were developed based on the batch equilibrium data. Based on these isotherms, the amount of activated coke required to achieve certain removal of color and AOX can be predicted. The utilization of the petroleum coke for the production of activated carbon can provide an excellent disposal option for the oil sand industry at the same time would provide a cheap and valuable activated carbon.

Charcoal↗

Lethal interaction of ubiquitous insecticide carriers with virus.

Large quantities of presumably nontoxic petroleum oil by-products are introduced into the environment as pesticide dispersal agents and emulsifiers. An increase in viral lethality with a concomitant influence on the liver and central nervous system occurs in young mice previously primed with such chemicals.

Animals↗

Specific acyclic isoprenoids as biological markers of methanogenic bacteria in marine sediments.

The widespread occurrence of extended hopanoids in sediments and petroleums illustrates the importance of bacterial lipid contributions to geological materials. In archaebacteria, however, hopanoids are absent; their role as structural components of biomembranes is fulfilled by acyclic isoprenoids. Recent studies of the lipid constituents of archaebacteria have greatly extended the range of acyclic isoprenoid skeletons known in organisms (Fig. 1). In particularly, isoprenoids with head-to-head linkages have been identified, and such compounds (for example, 3,7,11,15,18,22,26,30-octamethyldotriacontane, I) have been recognized in petroleum and as degradation products of Messel shale kerogen. Here we report the first recognition of 2,6,10,15,19-pentamethyleicosane (II), a known component of methanogens, in marine sediments of Recent to Cretaceous age (Table 1) and suggest that it and certain other acyclic isoprenoids may be used as biological markers for methanogens.

Euryarchaeota↗